Bijan Soleymani1, Shahram Parvaneh1, Ali Mostafaie1. 1. Medical Biology Research Center, Health Technology Institute, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Abstract
BACKGROUND: Sex selection of sperm by separating X- and Y-chromosome bearing spermatozoa is critical for efficiently obtaining the desired sex of animal offspring in the livestock industry. The purpose of this study was to produce a goat polyclonal antibody (pAb) against the bovine Sex Determining Region Y chromosome (bSRY) to separate female- and male-bearing spermatozoa. METHODS: To produce a goat polyclonal antibody against bSRY, a female goat was subcutaneously immunized with 27 kDa of recombinant bSRY (rbSRY) protein as the antigen. The anti-bSRY pAb was purified by ion-exchange chromatography. The purity of the pAb was determined using the SDS-PAGE method. The biological activity of the anti-bSRY pAb was examined using PCR to assess the binding affinity of pAb for the bSRY antigen and commercially sexed bull sperm. RESULTS: The total amount of purified anti-bSRY pAb was approximately 650 mg/goat serum (13 mg/mL). Interestingly, our data showed that the binding affinity of our pAb to the Y bearing was high, while the binding affinity of that to the X-chromosome bearing sperm was similar to the negative control. CONCLUSION: In conclusion, our findings show that the goat anti-SRY pAb specifically binds to Y-chromosome bearing sperm that suggesting its potential use for sex selection.
BACKGROUND: Sex selection of sperm by separating X- and Y-chromosome bearing spermatozoa is critical for efficiently obtaining the desired sex of animal offspring in the livestock industry. The purpose of this study was to produce a goat polyclonal antibody (pAb) against the bovine Sex Determining Region Y chromosome (bSRY) to separate female- and male-bearing spermatozoa. METHODS: To produce a goat polyclonal antibody against bSRY, a female goat was subcutaneously immunized with 27 kDa of recombinant bSRY (rbSRY) protein as the antigen. The anti-bSRY pAb was purified by ion-exchange chromatography. The purity of the pAb was determined using the SDS-PAGE method. The biological activity of the anti-bSRY pAb was examined using PCR to assess the binding affinity of pAb for the bSRY antigen and commercially sexed bull sperm. RESULTS: The total amount of purified anti-bSRY pAb was approximately 650 mg/goat serum (13 mg/mL). Interestingly, our data showed that the binding affinity of our pAb to the Y bearing was high, while the binding affinity of that to the X-chromosome bearing sperm was similar to the negative control. CONCLUSION: In conclusion, our findings show that the goat anti-SRY pAb specifically binds to Y-chromosome bearing sperm that suggesting its potential use for sex selection.
Entities:
Keywords:
Polyclonal antibody; Sex determining region Y chromosome; Sperm sexing
Authors: D Rath; S Barcikowski; S de Graaf; W Garrels; R Grossfeld; S Klein; W Knabe; C Knorr; W Kues; H Meyer; J Michl; G Moench-Tegeder; C Rehbock; U Taylor; S Washausen Journal: Reproduction Date: 2013-01-08 Impact factor: 3.906